Trade-offs in cotton pest management: Seed treatments suppress pests but reduce the abundance of natural enemies in the arthropod community

Melis Yalçin

doi:10.1371/journal.pone.0346422

. 2026 Apr 21;21(4):e0346422. doi: 10.1371/journal.pone.0346422

Trade-offs in cotton pest management: Seed treatments suppress pests but reduce the abundance of natural enemies in the arthropod community

Melis Yalçin ^1,^*

Editor: Rachid Bouharroud²

PMCID: PMC13098939 PMID: 42013091

Abstract

Pesticide seed treatments are widely used to protect crops from early-season pests and diseases; however, their broader ecological effects on arthropod communities and non-target organisms remain insufficiently understood. A two-year field experiment was conducted in a cotton agroecosystem to evaluate the community-level effects of seed-applied clothianidin (CLO) and a fungicide mixture containing azoxystrobin, metalaxyl-m, and fludioxonil (AMF). Changes in arthropod population density and diversity were assessed across taxonomic and community levels. The dominant arthropod families recorded included Nabidae, Miridae, Anthocoridae, Asilidae, Chrysopidae, Cicadellidae, and Coccinellidae. Both seed treatments suppressed key sucking pest groups, particularly Cicadellidae and Miridae; however, a significant reduction in predator abundance was also observed in treated plots. These findings indicate a potential trade-off between effective pest suppression and the conservation of natural enemy populations within the arthropod community. Principal response curve (PRC) analysis further demonstrated treatment-related shifts in arthropod communities over time. The results highlight the importance of considering non-target ecological responses when evaluating the sustainability of prophylactic seed treatments in cotton agroecosystems and emphasize the need for long-term investigations into their impacts on multitrophic interactions.

Introduction

Cotton (Gossypium hirsutum L.) is a significantly important cash crop grown in a hundred countries across the globe [1,2] and an indispensable natural textile fiber commodity, often called white gold, and the king of fiber [3]. It fulfills one-third of the natural fiber demand of the world population [4]. However, cotton production faces serious threats from diverse arthropod pests and diseases. The damage caused by soil-borne diseases, early-season sap-sucking insect pests, and other pests to cotton crops require sustainable pest management strategies to mitigate economic and crop production losses. Seed coating is one of the most crucial management strategies to protect seeds and seedlings. This technique was first used in 1930 for cereal seeds to prevent damage from soil-borne pathogens and became widespread between the 1970s and 1980s on field crops such as soybean, corn, wheat, and cotton [5,6].

Clothianidin (CLO) is a soil-active neonicotinoid insecticide commonly used for soil and plant pests. CLO targets nicotinic acetylcholine receptors (nAChR) and promotes paralysis and hyperstimulation of pests [7,8]. It is used to control black cutworm Agrotis ipsilon, wireworms from Coleoptera: Elateridae and Coleoptera: Tenebrionidae, White grubs, Coleoptera: Scarabaeidae, Cotton Flea beetle Coleoptera: Chrysomelidae, Homoptera: Aphididae, and Thysanoptera: Thripidae. Azoxystrobin is a synthetic, biodegradable, translaminar strobilurin fungicide [9]. As a mode of action, it can inhibit mitochondrial respiration between cytochromes [10]. It is generally combined with metalaxyl-m and fludioxonil. AMF is generally used on cotton for Rhizoctonia solani, Fusarium, and Phytium. The chemical properties of these pesticides affect their uptake and translocation in the plant. The lipophilicity, volatility, and polarity of the pesticide components and the vegetation period of the products are crucial for calculating the risk assessment and determining the amount of pesticides that are transferred through different parts of the plant [11–13].

Natural enemies may be exposed to insecticides and fungicides directly or indirectly through consumption of herbivores, contact with treated plant surfaces, or feeding on nectar, extrafloral nectar, pollen, honeydew, and guttation liquid [11,14–17]. Adverse effects have been reported for several taxa, including Coccinellidae, Anthocoridae, Nabidae, Chrysopidae, spiders, Braconidae, and Syrphidae [5,11,17–20]. Such exposure can reduce natural enemy abundance and cause non-target effects, particularly during the flowering period when nectar and pollen attract beneficial arthropods and guttation fluid provides water resources [21,22]. For example, Apis mellifera exposed to clothianidin contaminated guttation fluid from maize plants exhibited high mortality, indicating that systemic insecticides can affect non-target beneficial insects through plant-derived exposure routes [23]. Clothianidin seed treatments may also indirectly affect natural enemies by reducing prey availability, sometimes leading to pest outbreaks such as increased Tetranychus sp. populations [24]. Although some benefits have been reported, studies demonstrate that seed treatments with clothianidin, imidacloprid, or thiamethoxam can disrupt arthropod community structure, alter diversity and evenness, and decrease beneficial predator abundance [14,25]. Still, the effects of seed-applied pesticides on arthropod communities under field conditions are not fully understood.

This study examined how seed-applied pesticides influence sucking pest populations, a major issue in cotton agroecosystems, and their natural enemies at the community level. The abundance and diversity of both pests and predators were evaluated. Additionally, clothianidin (CLO) and a fungicide mixture (AMF) were applied as seed treatments to assess their efficacy and compare arthropod population densities across different pesticide applications. The findings provide new insights into the effects of seed treatments on cotton sucking pests and their potential side effects on predator communities.

Materials and methods

Study site

The study was conducted at the Aydın Adnan Menderes University Faculty of Agriculture Research and Application Farm (37°45′32″N, 27°45′35″E), Aydın, Turkey. Lodos cotton cultivar (Özaltın Seed Company of Turkey) was planted in the last week of May in 2021 and 2022, with 70 cm inter-rows and 15 cm intra-row spacing. The treatments were arranged in 20 and 10 m by a randomized complete block design with three replications. Each plot column was separated to ensure distinct boundaries and simplify sample collection; 1-meter bare strips were used to separate the columns. CLO was applied as a seed treatment (Poncho 600 Basf) at a rate of 800 ml 100 kg^-1 seed, and AMF was applied as a seed treatment (Dynasty CST 125 FS Syngenta) at a rate of 250 ml 100 kg^-1 seed, and also untreated seed as a control was used at the exact locations for each year. During the experiment, no pesticides were applied. Weeds were controlled with cultural practices (mainly tillage and hand-hoeing) during the growing season.

Experimental setup and arthropod collection methods

Foliar-sucking arthropods and their predators were sampled for twelve weeks between 18 June 2021 and 7 September 2021, and 17 June 2022 and 9 September 2022 until the cotton harvest time. The average physical conditions between seeding and harvest were found 0.02 mm and 1.1 mm of rainfall, 26.9 °C (max. 34.9 °C and min.17.8 °C), 26.3 °C (max. 33.5 °C and min.18.3 °C), and 53.33% R.H. (max. 89%, min. 33.6%), 66.44% R.H. (98.7%, min. 37.7%) (Meteorological Station, 2021–2022 Aydın, Turkey). Samples were collected 12 times during the cotton growing season because the most active time interval for pests and predators is from June to September. Arthropod abundance in the plant canopy was measured by sweep netting, where 50 sweeps were taken in a straight line through the center of each plot. Sweep net sampling was used to assess the abundance of mobile foliar-dwelling arthropods present in the plant canopy. Sweep netting was conducted at intervals of 100 meters from the field’s edge to avoid potential edge effects on the experimental results. Pests and predators were identified under a stereomicroscope (Leica E24, 10x) and recorded at the family level. Data from samples within three replicates were averaged for analysis. The interactions between the plant’s vegetative stage and pesticide treatment effects were observed for both years in 2021 and 2022 in the Aydın cotton province.

Statistical analysis

Arthropod diversity in the samples was assessed by identifying arthropods at the family level, with both adults and immature stages grouped for each taxon. Due to taxonomic uncertainty at the family level, the analyses did not include insects belonging to Diptera, Hymenoptera, and Lepidoptera.

The effects of seed treatment with CLO and AMF on the number of arthropods were determined using a general linear model (GLM). The sum of mean values of pests and natural enemies were compared using one-way ANOVA in SPSS, followed by Tukey’s HSD post hoc test.

The total number of taxonomic families (Taxa richness), number of individuals recorded (Total abundance), Shannon index, and evenness were calculated to quantify the diversity of the arthropod community in the treated and untreated plots for each week.

The Shannon-Wiener Diversity Index (H) is the most used diversity index in ecological experiments [26,27].

H = - \sum_{i = 1}^{s} P i ln P i

In the formula ‘ln’is the natural log, ‘pi’ is the proportion of individuals in the sum of the species.’∑’ is the sum of the results. ‘s’ is the total number of species.

Shannon Index and Evenness data were subjected to one-way ANOVA and compared using the LSD (least significant difference) test at the 0.05 probability level.

The overall composition of arthropod communities is based on each taxonomic group’s abundance relative to the total number of arthropods recorded. Principal Response Curve (PRC) analysis was employed to synthesize overall community-level responses of arthropods to seed treatments. Sample data was averaged for each plot, and a two-year data set was included in the same analysis. Experimental design and synchronized sampling based on cotton growth enabled valid comparisons across years. As key factors to explain potential changes for each community dataset, seed treatment and sampling time were used, and data were log(x) transformed to adjust the raw data to stabilize the effect of variance. Canonical coefficients were obtained in R (R Core Team, 2012) using the ‘vegan’ package [28,29] for each sampling date to track the arthropod community response to the seed treatment over time. These coefficients were plotted over time, revealing how the seed-treated community differed from the control group throughout the experiment. The arthropod group most affected by the seed treatment was computed according to taxon-specific weights. A Monte Carlo permutation was used to see if the treatment’s impact, captured by canonical coefficients, was indeed present at each time point. Four hundred ninety-nine new datasets were generated under the null hypothesis of no treatment effect while maintaining the observed treatment and sampling time structure. Sampling time, replication, and year-replication interactions were treated as covariates in the analysis, restricting data permutation to preserve repeated measures and blocking structure, effectively removing these effects from the residual error. The P-value determined the significance level, calculated as the proportion of F values from the permuted data greater than or equal to the observed F value.

Diversity measurements and abundances of individual or pooled taxa contributing significantly to the overall community response were calculated by the mean values and standard errors. Treatment effects were further evaluated with a mixed-model ANOVA, where seed treatment and sampling time were fixed effects, and replication and year were random effects, repeated measures adjusted for temporally correlated observations. Before analysis, residual plots and the Shapiro-Wilk’s W test were applied to assess data normality and homogeneity of variance assumptions. Transformations or variance grouping are used as needed to meet ANOVA assumptions.

Results

Effects on pest communities

The effects of insecticidal seed treatments on cotton fluctuated throughout the experiment, showing no consistent pattern (S1 Table). The interaction of the treatment and sampling weeks was changed for the arthropod groups. Across the years, the treatment effects on the combined population of Miridae and Cicadellidae were significant (p≤0.01). More abundant arthropod groups in the insecticide-treated plot included Cicadellidae and Miridae. The overall mean number of Cicadellidae per 50 sweep net for the first week was recorded as 844 ± 44.55 and 995 ± 12.90 in the untreated control plots in 2021 and 2022, respectively. Analysis of individual arthropod groups showed a significant reduction in the overall abundance of insecticide-treated plots. In 2021, the abundance of Cicadellidae exhibited marked fluctuations, particularly between the third and eighth weeks of observation during the vegetative and flowering and boll retention stages (S1 Fig). Notably, as Cicadellidae densities decreased, Miridae populations tended to increase, suggesting a possible compensatory dynamic between the two taxa (p < 0.05). Clothianidin seed treatment consistently reduced Cicadellidae abundance compared to the untreated control across all phenological stages. However, on six sampling dates 25 June (F = 5.903, p = 0.038), 1 July (F = 10.858, p = 0.010), 9 July (F = 7.869, p = 0.021), 19 July (F = 68.662, p < 0.001), 5 August (F = 58.644, p = 0.001), and 19 August (F = 68.458; p < 0.001) no significant differences were found between the clothianidin and azoxystrobin treatments during the flowering and boll development and opening stages.

In 2022, the Cicadellidae population was consistently more abundant in the untreated control cotton plot, particularly during the vegetative, flowering, and boll retention stages, compared to the CLO seed treatment. Differences between the control and CLO-treated plots were statistically significant throughout the season. However, on 24 June (F = 2.000, p = 0.216), 29 July (F = 2.558, p = 0.157), 12 August (F = 0.139, p = 0.873), and 26 August (F = 2.878, p = 0.133), corresponding to the flowering and boll development and opening stages, no significant differences were detected between the two treatments (S2 Fig).

The mean (±SEM) seasonal densities of Miridae per 50 sweep net for the first week in the untreated control plots were 55 ± 3.53 in 2021 and 93 ± 14.2 in 2022. CLO and AMF suppressed the Miridae population throughout the crop-growing season of 2021. Only on 19 August (F = 71.695, p < 0.001), AMF seed treatment was less effective than before. In the last two weeks of the 2021 cotton growth season, no significant change in population density was observed in the pesticide seed treatment plots compared to the untreated control plots. This may be related to the fact that the last two weeks coincide with the end of the season (S3 Fig).

In 2022, there weren’t any essential differences between the population of Miridae in the untreated control group and treated areas for the first (F = 2.160; p = 0.197) and last two weeks (F = 3.000; p = 0.125) and also in the 4 August (F = 1.807; p = 0.243) and 19 August (F = 1.623; p = 0.273) (S1 Table). CLO seed treatment diminished mean seasonal densities for seven weeks of the 2022 growing season. The lower Miridae population observed with the CLO seed treatment compared to the untreated control and AMF seed treatment indicates that CLO played a key role in reducing the pest population (S4 Fig).

Across the 2021 and 2022 growing seasons, the seasonal mean abundances of Miridae and Cicadellidae differed significantly among seed treatments (Table 1). For Miridae, treatment effects were highly significant in 2021 (F = 1871.29, p < 0.001) and remained significant in 2022 (F = 53.52, p = 0.0013). Seasonal comparisons indicated that Miridae abundance was consistently highest in control plots, lowest in clothianidin-treated plots, and moderate in AMF-treated plots across both years.

Table 1. Seasonal mean abundance of Miridae and Cicadellidae under different seed treatments in 2021 and 2022.

2021	Control	CLO	AMF	F	P
Miridae	257.67 ± 56.69 a	107.00 ± 23.71c	164.00 ± 30.20 b	1871.29	<0.001
Cicadellidae	577.67 ± 69.75 a	263.67 ± 40.60 c	409.67 ± 67.56 b	276.03	<0.001
2022
Miridae	231.00 ± 39.37 a	70.67 ± 14.97 c	149.00 ± 24.60 b	53.52	37.86
Cicadellidae	419.00 ± 53.43 a	192.33 ± 33.82 b	277.00 ± 40.86 b	0.0013	0.0025

Open in a new tab

Means with row followed by the same letter are not significantly different from each other according to Tukey’s HSD test, α = 0.05

Similarly, Cicadellidae abundance differed significantly among treatments in both years (F = 276.03, p < 0.001 in 2021; 2022: F = 37.86, p = 0.0025 in 2022). Seasonal patterns were comparable to those observed for Miridae, with control plots showing the highest abundances, clothianidin-treated plots the lowest, and AMF-treated plots moderate values in both growing seasons. Overall, these results demonstrate a consistent treatment effect across years, with clothianidin treatments associated with the greatest reduction in sucking pest populations (Table 1).

Effects on predator communities

CLO and AMF-seed-treated plots had significantly fewer predators than the untreated control plots. The total predator population was drastically reduced due to seed treatments in 2021. Field treated with clothianidin seed treatment showed a significant reduction in predator population by approximately 43%, from a mean value of 4.39± 0.38 predators per sweep net in untreated cotton to 2.48± 0.27 in treated fields. Similarly, fields treated with a combination of AMF decreased 26% in the total predator population from a mean value of 4.39± 0.38 predators per sweep net in untreated cotton to 3.22 ± 0.33 in treated cotton fields (F = 1.693, p = 0.225). The study again highlighted a considerable effect of seed treatment on the predator population in the following year, 2022. The seed treatment with clothianidin resulted in a 75% reduction in the predator population from a mean value of 3.38 ± 0.13 per sweep net in untreated cotton to 0.83± 0.06 per sweep net in treated fields. Meanwhile, the field treated with the AMF resulted in a 57% decline in the total predator population from a mean value of 3.38 ± 0.13 to 1.43 ± 0.06 predators per sweep net (F = 3.933, p = 0.049) (Table 2). The taxa that exhibited significant reductions in treated fields across both years included Chrysopidae, Nabidae, Coccinellidae, Anthocoridae, and Asilidae. During the flowering and boll retention stage for both years, the Anthocoridae population peaked at the beginning of August (S5 and S6 Fig). Among the treatments, CLO seed treatment resulted in lower population densities than the untreated control and showed significant differences between the control and AMF seed treatment (Table 2). The total abundance of the Asilidae community was higher in the untreated control group for both years, and in 2021, the CLO seed treatment led to low Asilidae population density compared to AMF seed treatments (F = 5.089, p = 0.008) (Table 2). During the different development stages, changes in the Asilidae population exhibited inconsistent variation, reaching its highest level in both years in August (S7 and S8 Fig). The total abundance of the Coccinellidae population did not significantly change under pesticide seed treatments in 2021 (F = 1.844, p = 0.163), but it was lower in 2022 (F = 15.386, p = 0.001) (Table 2). The abundance of the Coccinellidae population showed inconsistent variation in the development stages for both years. However, the population density peaked in July for both years (S9 and S10 Fig). The abundance of predatory Chrysopidae was reduced in both insecticide treatments compared to the untreated control in 2021 (F = 3.149, p = 0.001) and in 2022 (F = 26.654, p = 0.001) (Table 2). It is noteworthy that in both years, seed treatment had a similar variable impact on the abundance of predatory Chrysopidae, while the number of Chrysopidae population reduced in July and peaked between August in both years (S11and S12 Fig). Numbers of Nabidae were significantly lower with CLO and AMF seed treatments in 2021 (F = 4.761, p = 0.01) and in 2022 (F = 15.163, P = 0.00) (Table 2). According to the interactions between treatment and plant stage, the Nabidae population was reduced from the vegetative stage through the end of the flowering and boll retention stage in both years (S13and S14 Fig).

Table 2. Mean sums of cotton natural enemy population collected per plot using sweep nets.

2021	Control	CLO	AMF	F	P
Chrysopidae	3.50 ± 0.39a	2.19 ± 0.35b	2.92 ± 0.35ab	3.149	0.047
Nabidae	1.83 ± 0.27a	1.08 ± 0.146b	1.03 ± 0.17b	4.761	0.010
Coccinellidae	6.92 ± 1.33a	3.92 ± 0.84a	4.92 ± 1.12a	1.844	0.163
Anthocoridae	6.36 ± 1.02a	3.44 ± 0.93b	4.19 ± 0.99ab	2.396	0.096
Asilidae	3.36 ± 0.42a	1.78 ± 0.27b	3.03 ± 0.41a	5.089	0.008
Total Predators	21.97 ± 0.38a	12.41 ± 0.27b	16.09 ± 0.33b	1.693	0.225
2022
Chrysopidae	3.72 ± 0.81a	0.86 ± 0.47b	1.39 ± 0.39b	26.654	0.001
Nabidae	1.44 ± 0.36a	0.44 ± 0.28b	0.53 ± 0.28b	15.163	0.001
Coccinellidae	1 ± 0.26a	0.17 ± 0.14b	0.25 ± 0.16b	15.386	0.001
Anthocoridae	6.56 ± 0.89a	1.42 ± 0.36b	3.25 ± 0.55ab	2.396	0.096
Asilidae	4.19 ± 0.75a	1.28 ± 0.38b	1.75 ± 0.35b	21.974	0.001
Total Predators	16.91 ± 0.13a	4.17 ± 0.06b	7.17 ± 0.06ab	3.933	0.049

Open in a new tab

Means with row followed by the same letter are not significantly different from each other according to Tukey’s HSD test, α = 0.05

Community-level implications

The principal response curve shows the effects of CLO and AMF-treated cotton seed on the arthropod community for 12 weeks. Analysis was performed with sweep net capture data as plot averages pooled over both study sites. The first ordination axis explained 68.7% of the variance with sweep net data. It indicated that the overall community response in the insecticide-treated plots significantly deviated from the zero-reference line representing the control community (p≤0.01). Taxon weight indicates which arthropod groups contributed the most to the community response. Higher positive weight indicates that arthropod abundances in the insecticide-treated plots followed the trend depicted by the response curve, whereas higher negative values reveal the opposite. The most affected arthropod groups include Nabidae, Miridae, Anthocoridae, Asilidae, Chrysopidae, Cicadellidae, and Coccinellidae (Fig 1).

Principal response curve (PRC) showing the effects of seed treatments (clothianidin, CLO; azoxystrobin + metalaxyl-M + fludioxonil, AMF) on arthropod community composition over time during the cotton growing seasons. The untreated control is represented by the horizontal baseline (zero line), and treatment effects are expressed as deviations from the control across sampling weeks. Species weights indicate the contribution of each arthropod group to the observed community responses; taxa with higher absolute weights are more strongly associated with treatment effects.

The difference between years in terms of the effect of seed treatments on the Shannon-Wiener and Evenness Indexes of predators were significant. However, seed treatments and seed treatments × year interactions were found to have no significant effect on the Shannon-Wiener and Evenness Index of predators (Table 3). The Shannon-Wiener and Evenness Index were higher in 2021 by about 19.07% and 19.16%, respectively (Table 4).

Table 3. Results (F-values) of repeated-measures (weeks) ANOVA of the effects of seed treatments, years, and their interaction on the Shannon-Wiener and Evenness Index of predators.

Community Diversity Indexes	Effects	df	Predators
Shannon-Winer	Year (Y)	1	4.23*
	Seed Treatment (ST)	2	0.54
	Y × ST	2	0.11
Evenness	Year (Y)	1	4.22*
Evenness	Seed Treatment (ST)	2	0.54
	Y × ST	2	0.11

Open in a new tab

*: p ≤ 0.05, **: p ≤ 0.01, no asterisk: nonsignificant differences.

Table 4. The comparison of seed treatments regarding Shannon-Wiener and Evenness Index in each year separately.

Years	Treatments	Shannon-Wiener Index	Evenness Index
2021	Control	1.035	0.643
	CLO	0.938	0.583
	AMF	1.000	0.621
	Year Average	0.991 A	0.616 A
2022	Control	0.890	0.553
	CLO	0.763	0.474
	AMF	0.751	0.467
	Year Average	0.802 B	0.498 B

Open in a new tab

Means not followed by the same letter(s) in the same column are significantly different. Upper-case letters were used to compare years with each other.

Discussion

Effects of pesticides on pest communities

This experiment investigated the community-level effects of AMF and CLO on arthropods in the cotton agroecosystem. A total of 7 families were recorded in the foliar communities, showing variation among the seed treatments with CLO, AMF, and the untreated control. PRC analyses indicated changes in taxon abundances mainly during the vegetation period of cotton. Overall, in both years (2021 and 2022), the arthropods on cotton plants were more significantly affected by CLO than AMF treatment. Two herbivore taxa were prevalent on cotton in our two-year experiment, including Cicadellidae and Miridae, particularly during the vegetative, flowering, and boll development stages of cotton. These results confirm that soybean-based neonicotinoid seed treatments can significantly suppress early-season herbivore populations [17]. Similarly, imidacloprid-treated cotton has been shown to effectively reduce Amrasca devastans populations while causing minimal impact on certain natural enemies [30]. However, AMF treatment had inconsistent effects on pest populations. Especially in 2021, it resulted in moderate suppression of leafhoppers and mirids, though the effect was less evident than that of clothianidin. These findings are consistent with those of [5,31], who observed that fungicide seed treatments are not primarily used for insect control; they may indirectly alter arthropod communities by interacting with environmental conditions and predator dynamics. According to the experiments about pesticide seed treatment effects on arthropod community, herbivores, particularly thrips [25], soybean aphids [32], sugarcane aphids [20], and leafhoppers were negatively affected by clothianidin than other taxon groups, in contrast, some groups such as grasshoppers [17], Lygus sp., Elaterids [18] Collembolans [25] exhibited significantly higher abundances in the seed treated plots. Within the current study, the population dynamics of Cicadellidae highlighted considerable variation between the two years. In 2021, the peak densities of Cicadellidae were primarily observed during the vegetative and boll opening stages, particularly 115–126 days after sowing (DAS), whereas in 2022, the population was more concentrated in vegetative and anthesis stages (initial high densities around 44 DAS, later highest around 86 DAS). The year-to-year variation in pest population showed different environmental conditions in the two growing seasons in Aydın, potentially influencing population dynamics and pest phenology. The comparison of these findings is consistent with the study of Saeed et al. [30], which highlighted similarities in early population development. Saeed et al. [30] reported that Amrasca devastans (Hemiptera: Cicadellidae) population peaked at 50 and 55 days post-sowing (DAS) following imidacloprid and thiamethoxam treatment in cotton during 2010 and 2011, respectively, which corresponds closely with the early period of high Cicadellidae density observed around 44–52 DAS in the current study.

The seasonal peak of Miridae abundance, recorded between late July and early August (approximately 84–101 DAS in 2021 and 86–100 DAS in 2022) in the current study, is consistent with findings from other cotton-growing regions [33–35]. Notably, the absence of significant differences in the Miridae population among treatments during the final two weeks of sampling (beyond ~115 DAS) suggests that the residual efficacy of seed-applied insecticides, particularly CLO, had likely declined by the late boll development stage.

This observation is consistent with the anticipated degradation of systemic insecticides over a growing season and supports findings by Zhang et al. [36], who also reported reduced efficacy of nitenpyram seed treatment against mirid bug Apolygus lucorum during later sampling periods.

The variation observed in the current study about the effectiveness of CLO compared to the consistent early-season control could be attributed to differences in the dominant species present in the community, regional environmental factors affecting systemic uptake and persistence, or local agroecological conditions in the region. Therefore, while the population of Cicadellidae and Miridae indicated predictable seasonal peaks, their susceptibility to specific seed treatments like CLO may be variable and suggests further evaluation, explicitly concerning the dominant species and the duration of effective suppression under diverse regional conditions.

Effects of pesticides on predator communities

The study shows that pesticide seed treatments significantly reduce predatory insect populations, consistent with previous research [37–42]. These predators are essential for regulating pest outbreaks in agroecosystems [14,25,32,43]. Systemic pesticides may disrupt predator communities, weakening natural pest control and increasing pesticide dependence [5,18]. In this study, the neonicotinoid insecticide CLO and the fungicide mixture AMF treatments reduced predator densities over two years, indicating effective translocation within the plant. The movement of systemic pesticides depends on properties such as lipophilicity and log Kow, which enhance their solubility and distribution to various plant parts [11]. This systemic spread increases the risk of exposure to non-target organisms. Furthermore, previous studies emphasize the vulnerability of omnivorous predators such as Geocoris, Orius, and Coccinellids due to their feeding behaviour, such as consuming nectar, pollen, plant tissues, and herbivorous prey; these natural enemies are exposed to both direct and indirect pathways of pesticide uptake. For example, indirect poisoning can occur when predators consume herbivores like leafhoppers that have ingested neonicotinoids [10,30,43,44]. Additionally Lysiphlebus testaceipes (Cresson) (Hymenoptera: Braconidae) populations were significantly reduced after exposure to thiamethoxam contaminated extrafloral nectar [16].

Particularly during the early vegetative stage of the plant, the higher mortality of natural enemies can be explained by two factors: the intense residue levels due to the plant’s vegetative stage and the fact that predators are in their immature stages, during which they have a greater need for plant nutrients [11,45,46]. According to the experiment of Gontijo et al. [11], the insecticidal toxicity of thiamethoxam and clothianidin was diminished with plant development, and thiamethoxam-coated seed caused lethal and sublethal effects on O. insidiosus (Hemiptera: Anthocoridae) while clothianidin-coated seed led to sublethal impacts. In a study conducted on imidacloprid and thiamethoxam treatment on soybean seed, the general predator community was reduced by 25%, and similar predators, with the present study, such as O. insidiosus, Nabis americoferus (Hemiptera: Nabidae), and Chrysoperla (Neuroptera: Chrysopidae) populations were reduced in the pesticide-treated plots [17]. The current study results coincide with previous findings and demonstrate even more significant reductions in predator abundance, 44% in 2021 and 75% in 2022. These reductions surpass the 34.5% decrease observed in foliage communities within clothianidin-treated corn agroecosystems [25] and the 16% reduction reported in a meta-analysis by Douglas and Tooker [5].

Effects of pesticides on arthropod communities

Pesticides reduced the pest population in cotton crops in the experiment, and the lack of pest population in sunflower, corn, and soybean was detected in the other studies [14, 17,18,32]. Pests’ responses to pesticide treatment differ according to insect abundance and diversity [18,47].

Using PRC analysis, the current study evaluated the community-level effects of CLO and AMF seed treatments on arthropods in an Aydın cotton agroecosystem. This study provides novel field-based evidence that seed-applied systemic pesticides can influence not only target sucking pest populations but also the abundance and diversity of associated predator communities in cotton agroecosystems. By simultaneously examining pest and predator assemblages, the present work contributes to a more comprehensive understanding of how prophylactic seed treatments shape multitrophic arthropod interactions under field conditions. According to the PRC results, certain predatory insect families—most notably Coccinellidae, Chrysopidae, Asilidae, Anthocoridae, and Nabidae—as well as pest groups such as Cicadellidae and Miridae, exhibited significant negative responses to the pesticide in terms of overall predator species richness. However, the difference in predator diversity between years suggests that factors such as seed treatments, cropping practices, prey availability, or other abiotic influences such as climate variability may have negatively impacted predator communities [48]. Additionally, the evenness index in the present study declined from 0.616 in 2021 to 0.498 in 2022, indicating a reduction in the balance of species distribution over time. This finding contrasts with the higher evenness values reported by Firake and Behere [27] and Disque et al. [25], aligning with prior research suggesting that seed-applied insecticides can adversely impact the predator community. Consistent with previous studies, clothianidin treatments affected predator abundance more than overall diversity [5,25,44]. Predator diversity and evenness significantly decreased from 2021 to 2022, with untreated control plots consistently exhibiting the highest diversity and evenness values, showing the disruptive effect of seed-applied pesticides on predator communities. This study’s Shannon-Wiener diversity index values ranged from 0.751 to 1.035, with annual means of 0.991 in 2021 and 0.802 in 2022. These values were considerably lower than those reported by Firake and Behere [27] in northeast India, where Shannon index values typically exceeded 2.16, reflecting higher predator diversity in maize agroecosystems. Overall, these findings suggest that while seed-applied pesticides may not drastically reduce predator species richness at the community level, they can suppress predator abundance and disrupt the evenness of arthropod predator communities. From an agroecosystem sustainability perspective, these results highlight the importance of moving beyond target pest suppression when evaluating the ecological consequences of seed-applied systemic pesticides in cotton production systems. These findings provide an ecological basis for improving integrated pest management strategies by emphasizing the importance of balancing effective pest suppression with the conservation of beneficial predator communities in cotton agroecosystems.

Conclusion

Pesticide seed treatments are commonly applied in fields without consistent pest pressure as a precaution against pest outbreaks. CLO and AMF seed treatments significantly reduced arthropod populations in the Aydın cotton agroecosystem, with communities recovering over the growing season. The reduction in the Cicadellidae population leads to an increased Miridae population. Despite impacts on pest abundance, the pesticide seed treatment reduced the natural enemy population. The Shannon-Wiener diversity index and evenness of predators were analyzed for two consecutive years. The results showed no significant effect of CLO and AMF seed treatment on community diversity and evenness, but diversity and evenness were substantial over the years. The difference between years may be cropping practices, prey availability, or other abiotic influences, such as climate variability, that may have negatively impacted predator communities. According to the principal response curve analyses, the most affected arthropod group from seed treatment was observed: Nabidae, Miridae, Anthocoridae, Asilidae, Chrysopidae, Cicadellidae, and Coccinellidae. These findings highlight the importance of continuously and systematically monitoring pest and predator populations at the community level in cotton agroecosystems. Future research integrating species-level identification, longer-term field monitoring, and assessments of sublethal or behavioural effects on natural enemies would further improve understanding of the ecological consequences of prophylactic seed treatments and their implications for multitrophic interactions and biological control within sustainable IPM-based cotton production systems.

Supporting information

S1 Table. Mean sums of Cicadellidae and Miridae populations collected per plot using sweep nets.

(PDF)

pone.0346422.s001.pdf^{(149.5KB, pdf)}

S2 Table. Mean (± SD) abundance of arthropod families (Miridae, Cicadellidae, Anthocoridae, Asilidae, Coccinellidae, and Nabidae) under different seed treatments across sampling weeks in 2021 and 2022.

(PDF)

pone.0346422.s002.pdf^{(198KB, pdf)}

S1 Fig. Impact of pesticide treatments on Cicadellidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s003.pdf^{(75KB, pdf)}

S2 Fig. Impact of pesticide treatments on Cicadellidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s004.pdf^{(76.4KB, pdf)}

S3 Fig. Impact of pesticide treatments on Miridae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s005.pdf^{(73.3KB, pdf)}

S4 Fig. Impact of pesticide treatments on Miridae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s006.pdf^{(71.3KB, pdf)}

S5 Fig. Impact of pesticide treatments on Anthocoridae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s007.pdf^{(94.7KB, pdf)}

S6 Fig. Impact of pesticide treatments on Anthocoridae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s008.pdf^{(103.2KB, pdf)}

S7 Fig. Impact of pesticide treatments on Asillidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s009.pdf^{(93.4KB, pdf)}

S8 Fig. Impact of pesticide treatments on Asillidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s010.pdf^{(118.1KB, pdf)}

S9 Fig. Impact of pesticide treatments on Coccinellidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s011.pdf^{(113.2KB, pdf)}

S10 Fig. Impact of pesticide treatments on Coccinellidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s012.pdf^{(113KB, pdf)}

S11 Fig. Impact of pesticide treatments on Chrysopidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s013.pdf^{(110.5KB, pdf)}

S12 Fig. Impact of pesticide treatments on Chrysopidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s014.pdf^{(98.7KB, pdf)}

S13 Fig. Impact of pesticide treatments on Nabidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s015.pdf^{(110.7KB, pdf)}

S14 Fig. Impact of pesticide treatments on Nabidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s016.pdf^{(95.2KB, pdf)}

Data Availability

All relevant data are within the manuscript and its Supporting Information files.

Funding Statement

The author(s) received no specific funding for this work.

References

1.Ünay A, Balcï S, Cïnar VM. Gene action and combining ability for neps and seed coat neps in cotton (Gossypium hirsutum L.). Turkish J Field Crops. 2019;24(2):211–4. doi: 10.17557/tjfc.568326 [DOI] [Google Scholar]
2.Jabran K, Chauhan BS. Cotton production. Hoboken (NJ): John Wiley & Sons; 2019. doi: 10.1002/9781119385523 [DOI] [Google Scholar]
3.Naveed M, Maqsood MF, Cheema AR. Modeling the contribution of district-level cotton production to aggregate cotton production in Punjab (Pakistan): an empirical evidence using correlated component regression approach. J Excell Soc Sci. 2024;3(3):147–64. doi: 10.69565/jess.v3i3.200 [DOI] [Google Scholar]
4.Mollaee M, Mobli A, Mutti NK, Manalil S, Chauhan BS. Challenges and opportunities in cotton production. cotton production. Wiley; 2019. 371–90. doi: 10.1002/9781119385523.ch18 [DOI] [Google Scholar]
5.Douglas MR, Tooker JF. Meta-analysis reveals that seed-applied neonicotinoids and pyrethroids have similar negative effects on abundance of arthropod natural enemies. PeerJ. 2016;4:e2776. doi: 10.7717/peerj.2776 [DOI] [PMC free article] [PubMed] [Google Scholar]
6.Smalling KL, Hladik ML, Sanders CJ, Kuivila KM. Leaching and sorption of neonicotinoid insecticides and fungicides from seed coatings. J Environ Sci Health B. 2018;53(3):176–83. doi: 10.1080/03601234.2017.1405619 [DOI] [PubMed] [Google Scholar]
7.Bandeira FO, Alves PRL, Hennig TB, Brancalione J, Nogueira DJ, Matias WG. Chronic effects of clothianidin to non-target soil invertebrates: ecological risk assessment using the species sensitivity distribution (SSD) approach. J Hazard Mater. 2021;419:126491. doi: 10.1016/j.jhazmat.2021.126491 [DOI] [PubMed] [Google Scholar]
8.Hilton MJ, Emburey SN, Edwards PA, Dougan C, Ricketts DC. The route and rate of thiamethoxam soil degradation in laboratory and outdoor incubated tests, and field studies following seed treatments or spray application. Pest Manag Sci. 2019;75(1):63–78. doi: 10.1002/ps.5168 [DOI] [PMC free article] [PubMed] [Google Scholar]
9.Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M, Parr-Dobrzanski B. The strobilurin fungicides. Pest Manag Sci. 2002;58(7):649–62. doi: 10.1002/ps.520 [DOI] [PubMed] [Google Scholar]
10.Moser SE, Obrycki JJ. Non-target effects of neonicotinoid seed treatments; mortality of coccinellid larvae related to zoophytophagy. Biological Control. 2009;51(3):487–92. doi: 10.1016/j.biocontrol.2009.09.001 [DOI] [Google Scholar]
11.Gontijo PC, Moscardini VF, Michaud JP, Carvalho GA. Non-target effects of two sunflower seed treatments on Orius insidiosus (Hemiptera: Anthocoridae). Pest Manag Sci. 2015;71(4):515–22. doi: 10.1002/ps.3798 [DOI] [PubMed] [Google Scholar]
12.Özşirvan G, Yalçın M, Turgut N, Tari V, Turgut C. Insights into the uptake, translocation, and accumulation dynamics of cyantraniliprole and thiamethoxam seed coating pesticides in maize plants. Environ Sci Pollut Res Int. 2024;31(32):44900–7. doi: 10.1007/s11356-024-34135-7 [DOI] [PubMed] [Google Scholar]
13.Rein A, Trapp S, Fantke P, Yalçın M, Turgut N, Ahat C, et al. Uptake and translocation of pesticides in pepper and tomato plants. Pest Manag Sci. 2025;81(3):1562–70. doi: 10.1002/ps.8556 [DOI] [PMC free article] [PubMed] [Google Scholar]
14.Dubey A, Lewis MT, Dively GP, Hamby KA. Ecological impacts of pesticide seed treatments on arthropod communities in a grain crop rotation. J Appl Ecol. 2020;57(5):936–51. doi: 10.1111/1365-2664.13595 [DOI] [Google Scholar]
15.Yalçin M, Kapiz I, Kösoglu M, Koca YO, Turgut N, Topal E, et al. Translocation of clothianidin to guttation fluid and its potential impact on honey bee, Apis mellifera anatoliaca Maa, 1953 (Hymenoptera: Apidae). Turk Entomol Derg. 2021;45(4):511–22. doi: 10.16970/entoted.1017479 [DOI] [Google Scholar]
16.Moscardini VF, Gontijo PC, Michaud JP, Carvalho GA. Sublethal effects of insecticide seed treatments on two nearctic lady beetles (Coleoptera: Coccinellidae). Ecotoxicology. 2015;24(5):1152–61. doi: 10.1007/s10646-015-1462-4 [DOI] [PubMed] [Google Scholar]
17.Seagraves MP, Lundgren JG. Effects of neonicitinoid seed treatments on soybean aphid and its natural enemies. J Pest Sci. 2011;85(1):125–32. doi: 10.1007/s10340-011-0374-1 [DOI] [Google Scholar]
18.Bredeson MM, Lundgren JG. Thiamethoxam seed treatments have no impact on pest numbers or yield in cultivated sunflowers. J Econ Entomol. 2015;108(6):2665–71. doi: 10.1093/jee/tov249 [DOI] [PubMed] [Google Scholar]
19.Jones AG, Hoover K, Pearsons K, Tooker JF, Felton GW. Potential impacts of translocation of neonicotinoid insecticides to cotton (Gossypium hirsutum (Malvales: Malvaceae)) extrafloral nectar on parasitoids. Environ Entomol. 2020;49(1):159–68. doi: 10.1093/ee/nvz157 [DOI] [PubMed] [Google Scholar]
20.Szczepaniec A. Interactive effects of crop variety, insecticide seed treatment, and planting date on population dynamics of sugarcane aphid (Melanaphis sacchari) and their predators in late-colonized sorghum. Crop Protection. 2018;109:72–9. doi: 10.1016/j.cropro.2018.03.002 [DOI] [Google Scholar]
21.Pemberton RW, Lee JH. The influence of extrafloral nectaries on parasitism of an insect herbivore. Am J Bot. 1996;83(9):1187–94. doi: 10.1002/j.1537-2197.1996.tb13900.x [DOI] [Google Scholar]
22.Nicolson SW, Massimo N, Pacini E. Nectaries and nectar. Nicolson SW, Nepi M, Pacini E, editors. 2007. doi: 10.1007/978-1-4020-5937-7 [DOI] [Google Scholar]
23.Gao A-H, Fu Y-Y, Zhang K-Z, Zhang M, Jiang H-W, Fan L-X, et al. Azoxystrobin, a mitochondrial complex III Qo site inhibitor, exerts beneficial metabolic effects in vivo and in vitro. Biochim Biophys Acta. 2014;1840(7):2212–21. doi: 10.1016/j.bbagen.2014.04.002 [DOI] [PubMed] [Google Scholar]
24.Smith JF, Catchot AL, Musser FR, Gore J. Effects of aldicarb and neonicotinoid seed treatments on twospotted spider mite on cotton. J Econ Entomol. 2013;106(2):807–15. doi: 10.1603/ec10125 [DOI] [PubMed] [Google Scholar]
25.Disque HH, Hamby KA, Dubey A, Taylor C, Dively GP. Effects of clothianidin-treated seed on the arthropod community in a mid-Atlantic no-till corn agroecosystem. Pest Manag Sci. 2019;75(4):969–78. doi: 10.1002/ps.5201 [DOI] [PubMed] [Google Scholar]
26.Shannon CE. A Mathematical theory of communication. Bell System Tech J. 1948;27(3):379–423. doi: 10.1002/j.1538-7305.1948.tb01338.x [DOI] [Google Scholar]
27.Firake DM, Behere GT. Natural mortality of invasive fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) in maize agroecosystems of northeast India. Bio Control. 2020;148:104303. doi: 10.1016/j.biocontrol.2020.104303 [DOI] [Google Scholar]
28.Oksanen J, Blanchet FG, Minchin PR. Vegan: community ecology package. [Google Scholar]
29.Alberdi A, Gilbert MTP. Hilldiv: an R package for the integral analysis of diversity based on Hill numbers. 2019. http://biorxiv.org/lookup/doi/10.1101/545665
30.Saeed R, Razaq M, Hardy IC. Impact of neonicotinoid seed treatment of cotton on the cotton leafhopper, Amrasca devastans (Hemiptera: Cicadellidae), and its natural enemies. Pest Manag Sci. 2016;72(6):1260–7. doi: 10.1002/ps.4146 [DOI] [PubMed] [Google Scholar]
31.Mourtzinis S, Krupke CH, Esker PD, Varenhorst A, Arneson NJ, Bradley CA, et al. Neonicotinoid seed treatments of soybean provide negligible benefits to US farmers. Sci Rep. 2019;9(1):11207. doi: 10.1038/s41598-019-47442-8 [DOI] [PMC free article] [PubMed] [Google Scholar]
32.Regan K, Ordosch D, Glover KD, Tilmon KJ, Szczepaniec A. Effects of a pyrethroid and two neonicotinoid insecticides on population dynamics of key pests of soybean and abundance of their natural enemies. Crop Protection. 2017;98:24–32. doi: 10.1016/j.cropro.2017.03.004 [DOI] [Google Scholar]
33.Özgür O, Atakan E, Pehlïvan S. Investigation of the damage of Miridae species on cotton in Çukurova Region of Turkey. Turk Entomol Derg. 2019;43(2):143–56. doi: 10.16970/entoted.517428 [DOI] [Google Scholar]
34.Lu Y, Wu K, Jiang Y, Xia B, Li P, Feng H, et al. Mirid bug outbreaks in multiple crops correlated with wide-scale adoption of Bt cotton in China. Science. 2010;328(5982):1151–4. doi: 10.1126/science.1187881 [DOI] [PubMed] [Google Scholar]
35.Jiao Z, Jaworski CC, Lu Y, Ye L, Wu K, Desneux N. Maize fields are a potential sink for an outbreaking mirid bug pest in Chinese Bt-cotton agricultural landscapes. Agric Ecosyst Environ. 2019;279:122–9. doi: 10.1016/j.agee.2019.04.010 [DOI] [Google Scholar]
36.Zhang Z, Wang Y, Zhao Y, Li B, Lin J, Zhang X, et al. Nitenpyram seed treatment effectively controls against the mirid bug Apolygus lucorum in cotton seedlings. Sci Rep. 2017;7(1):8573. doi: 10.1038/s41598-017-09251-9 [DOI] [PMC free article] [PubMed] [Google Scholar]
37.Delbeke E, Vercruysse P, Tirry L, De Clercq P, Degheele D. Toxicity of diflubenzuron, pyriproxyfen, imidacloprid and diafenthiuron to the predatory bug Orius laevigatus (Het: Anthacoridae). Entomophaga. 1997;42(3):349–58. doi: 10.1007/BF02769828 [DOI] [Google Scholar]
38.Desneux N, Decourtye A, Delpuech J-M. The sublethal effects of pesticides on beneficial arthropods. Annu Rev Entomol. 2007;52:81–106. doi: 10.1146/annurev.ento.52.110405.091440 [DOI] [PubMed] [Google Scholar]
39.Papachristos DP, Milonas PG. Adverse effects of soil applied insecticides on the predatory coccinellid Hippodamia undecimnotata (Coleoptera: Coccinellidae). Biological Control. 2008;47(1):77–81. doi: 10.1016/j.biocontrol.2008.06.009 [DOI] [Google Scholar]
40.Peck DC. Long-term effects of imidacloprid on the abundance of surface- and soil-active nontarget fauna in turf. Agric For Entomol. 2009;11(4):405–19. doi: 10.1111/j.1461-9563.2009.00454.x [DOI] [Google Scholar]
41.Yao F-L, Zheng Y, Zhao J-W, Desneux N, He Y-X, Weng Q-Y. Lethal and sublethal effects of thiamethoxam on the whitefly predator Serangium japonicum (Coleoptera: Coccinellidae) through different exposure routes. Chemosphere. 2015;128:49–55. doi: 10.1016/j.chemosphere.2015.01.010 [DOI] [PubMed] [Google Scholar]
42.Tappert L, Pokorny T, Hofferberth J, Ruther J. Sublethal doses of imidacloprid disrupt sexual communication and host finding in a parasitoid wasp. Sci Rep. 2017;7:42756. doi: 10.1038/srep42756 [DOI] [PMC free article] [PubMed] [Google Scholar]
43.Albajes R, López C, Pons X. Predatory fauna in cornfields and response to imidacloprid seed treatment. J Econ Entomol. 2003;96(6):1805–13. doi: 10.1093/jee/96.6.1805 [DOI] [PubMed] [Google Scholar]
44.Pisa LW, Amaral-Rogers V, Belzunces LP, Bonmatin JM, Downs CA, Goulson D, et al. Effects of neonicotinoids and fipronil on non-target invertebrates. Environ Sci Pollut Res Int. 2015;22(1):68–102. doi: 10.1007/s11356-014-3471-x [DOI] [PMC free article] [PubMed] [Google Scholar]
45.Feese H, Wilde G. Planting time applications of systemic insecticides on grain sorghum for greenbug control: Interactions with herbicides and effect on predators (Hemiptera (Homoptera)). J Kans Entomol Soc. 1975;48(3):396–402. [Google Scholar]
46.Al-Deeb MA, Wilde GE, Zhu KY. Effect of insecticides used in corn, sorghum, and alfalfa on the predator Orius insidiosus (Hemiptera: Anthocoridae). J Econ Entomol. 2001;94(6):1353–60. doi: 10.1603/0022-0493-94.6.1353 [DOI] [PubMed] [Google Scholar]
47.Lundgren JG, Fausti SW. Trading biodiversity for pest problems. Sci Adv. 2015;1(6):e1500558. doi: 10.1126/sciadv.1500558 [DOI] [PMC free article] [PubMed] [Google Scholar]
48.Schneider CW, Tautz J, Grünewald B, Fuchs S. RFID tracking of sublethal effects of two neonicotinoid insecticides on the foraging behavior of Apis mellifera. PLoS One. 2012;7(1):e30023. doi: 10.1371/journal.pone.0030023 [DOI] [PMC free article] [PubMed] [Google Scholar]

PLoS One. doi: 10.1371/journal.pone.0346422.r001

Decision Letter 0

Rachid Bouharroud

4 Feb 2026

-->PONE-D-25-50468-->-->Balancing pest control and natural enemy conservation: Seed Treatment Effects in Cotton-->-->PLOS One

Dear Dr. Yalçın,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

-->-->When revising your manuscript and as requested by reviewer 2, please give special consideration to (Abstract, supporting materials, double check figures/tables references and discussion flow). Also you should well describe in detail the sweeping methods (trapped insects and stages) in Mat&Met section.-->--> -->-->Good luck-->-->

Please submit your revised manuscript by Mar 21 2026 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.. When you're ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

Please include the following items when submitting your revised manuscript:-->

A letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.
A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at . Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols....

We look forward to receiving your revised manuscript.

Kind regards,

Rachid Bouharroud

Academic Editor

PLOS One

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. Please ensure that you include a title page within your main document. We do appreciate that you have a title page document uploaded as a separate file, however, as per our author guidelines (httpp://journals.plos.org/plosone/s/submission-guidelines#loc-title-page) we do require this to be part of the manuscript file itself and not uploaded separately.

Could you therefore please include the title page into the beginning of your manuscript file itself, listing all authors and affiliations.

3. Please ensure that you include a title page within your main document. You should list all authors and all affiliations as per our author instructions and clearly indicate the corresponding author.

4. Please include captions for your Supporting Information files at the end of your manuscript, and update any in-text citations to match accordingly. Please see our Supporting Information guidelines for more information: http://journals.plos.org/plosone/s/supporting-information....

5. If the reviewer comments include a recommendation to cite specific previously published works, please review and evaluate these publications to determine whether they are relevant and should be cited. There is no requirement to cite these works unless the editor has indicated otherwise.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

-->Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. -->

Reviewer #1: Partly

Reviewer #2: Yes

**********

-->2. Has the statistical analysis been performed appropriately and rigorously? -->

Reviewer #1: I Don't Know

Reviewer #2: Yes

**********

-->3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.-->

Reviewer #1: Yes

Reviewer #2: Yes

**********

-->4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.-->

Reviewer #1: No

Reviewer #2: Yes

**********

-->5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)-->

Reviewer #1: Introduction is fairly long. A number of studies are cited, indicating a lot of literature already available on this topic. What are objectives of this study needs to be enlisted and introduction should be shaped according to them to build argument by identifying gaps rather than reviewing earlier literature.

L102: For the experimental design, three replicates were run following a randomized complete block design. In a small-scale study of this kind, different field as replicates may be more appropriate to include variation as true replicates.

L119: Sweep nests were used to assess abundance. If this method was used against flying adults or also when larvae or immobile or less immobile stages were present, needs clarity on this. how much time was spent per sweep.

Results: Seasonal counts can be compared among treatments at the end of season. Line graphs can be made supplementary and give their seasonal means comparison, results are too long and hard to follow under current data representation. Also provide information on dominant species of pests and predators.

Discussion: What authors find new in this study and discuss the way forward. What are the limitation of this study and possible research direction?

Reviewer #2: This two-year field study provides a valuable community-level assessment of the impact of two common seed treatments—clothianidin (CLO, insecticide) and azoxystrobin + metalaxyl-m + fludioxonil (AMF, fungicide combination)—on arthropod pests and their natural enemies in a cotton agroecosystem. The research addresses an important gap by moving beyond single-species assessments to evaluate whole-community dynamics, which is crucial for developing sustainable Integrated Pest Management (IPM) strategies.

Key Strengths of the Manuscript:

Strong Experimental Design: A two-year, replicated field trial with an untreated control provides robust, ecologically relevant data.

Comprehensive Community Analysis: The use of Principal Response Curve (PRC) analysis to synthesize community-level effects is a significant methodological strength, clearly visualizing how entire arthropod assemblages shift in response to treatments over time.

Important Findings: The study convincingly demonstrates that both CLO and AMF seed treatments significantly reduce the abundance of key natural enemy families (e.g., Nabidae, Chrysopidae, Coccinellidae) while also suppressing pest populations like Cicadellidae. The documented negative correlation between pest and predator suppression highlights a critical trade-off in pest management.

Clear Conclusions: The conclusions are well-supported by the data, emphasizing that seed treatments are not "safe" for natural enemies and calling for more judicious, need-based use within IPM frameworks.

Suggestions for Revision and Clarification:

Title Refinement: The current title ("Balancing pest control and natural enemy conservation: Seed Treatment Effects in Cotton") is slightly generic. Consider making it more specific to your key finding, e.g., "Trade-offs in cotton pest management: Seed treatments suppress pests but reduce natural enemy abundance in the arthropod community."

Abstract Enhancement: The abstract should more clearly state the main takeaway. Explicitly mention that your study found a significant reduction in predator abundance alongside pest suppression, creating a potential trade-off. Briefly specify the two treatments used.

Results Presentation: The text frequently refers to figures and tables (e.g., Fig 1, S1 Table) that are not included in the provided text file. Ensure all in-text citations have corresponding, clearly labeled items in the submitted manuscript.

Discussion Flow: The discussion is comprehensive but could be more tightly structured. Consider subheadings (e.g., "Effects on Pest Communities," "Effects on Predator Communities," "Community-Level Implications") to improve readability and guide the reader through your interpretations.

Clarity on "Neonicotinoid" Terminology: Clothianidin is a neonicotinoid. The introduction and discussion mention neonicotinoids broadly but could more explicitly frame CLO within this well-known class from the outset, as their non-target effects are a major point of discussion.

Statistical Reporting Consistency: Ensure that all statistical results (F-values, degrees of freedom, p-values) are reported consistently (e.g., F(1, 2) = 5.90, p = 0.038).

Data Availability Statement: The statement "The data supporting this study's findings are available in this article's supplementary material" is good, but PLOS ONE requires that the data be fully available without restriction. Double-check that all raw data underlying the figures and analyses are indeed included in the submitted Supporting Information files.

**********

-->6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.). If published, this will include your full peer review and any attached files.). If published, this will include your full peer review and any attached files.). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our For information about this choice, including consent withdrawal, please see our For information about this choice, including consent withdrawal, please see our For information about this choice, including consent withdrawal, please see our Privacy Policy..-->..-->

Reviewer #1: No

Reviewer #2: No

**********

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.]

To ensure your figures meet our technical requirements, please review our figure guidelines: https://journals.plos.org/plosone/s/figures

You may also use PLOS’s free figure tool, NAAS, to help you prepare publication quality figures: https://journals.plos.org/plosone/s/figures#loc-tools-for-figure-preparation.

NAAS will assess whether your figures meet our technical requirements by comparing each figure against our figure specifications.

PLoS One. 2026 Apr 21;21(4):e0346422. doi: 10.1371/journal.pone.0346422.r002

Author response to Decision Letter 1

18 Mar 2026

Answers to the reviewers comments

View Letter

Date: Feb 04 2026 12:16AM

To: "Melis Yalçın" melisusluy@gmail.com

From: "PLOS ONE" plosone@plos.org

Subject: PLOS ONE Decision: Revision required [PONE-D-25-50468]

PONE-D-25-50468

Balancing pest control and natural enemy conservation: Seed Treatment Effects in Cotton

PLOS One

Dear Dr. Yalçın,

When revising your manuscript and as requested by reviewer 2, please give special consideration to (Abstract, supporting materials, double check figures/tables references and discussion flow). Also you should well describe in detail the sweeping methods (trapped insects and stages) in Mat&Met section.

Good luck

Please include the following items when submitting your revised manuscript:

• A letter that responds to each point raised by the academic editor and reviewer(s). You should upload this letter as a separate file labeled 'Response to Reviewers'.

• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.

• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

Response to Reviewer: Thank you for your suggestion regarding the deposition of laboratory protocols in protocols.io. I appreciate the opportunity to enhance methodological transparency and reproducibility.

However, as the procedures applied in my study follow established and previously published methodologies, I believe that additional protocol deposition is not necessary at this stage.

We look forward to receiving your revised manuscript.

Kind regards,

Rachid Bouharroud

Academic Editor

PLOS One

Journal Requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

Response to Reviewer: Thank you for your reminder regarding compliance with PLOS ONE’s style requirements. I confirm that the revised manuscript has been carefully checked and formatted in accordance with the journal’s guidelines, including file naming conventions and the use of the appropriate PLOS ONE style template.

Could you therefore please include the title page into the beginning of your manuscript file itself, listing all authors and affiliations.

Response to Reviewer: Thank you for your clarification regarding the title page requirements. I have revised the submission accordingly and incorporated the title page at the beginning of the main manuscript file, including the author name and affiliation, in accordance with the PLOS ONE author guidelines. The manuscript has been re-uploaded with this modification.

Response to Reviewer: I would like to confirm that the title page has been prepared in accordance with the journal’s author guidelines. Author information and affiliation are included, and the corresponding author has been clearly indicated within the manuscript file.

Response to Reviewer: Thank you for your helpful comment regarding the Supporting Information files. In accordance with the journal’s guidelines, captions for all Supporting Information files have now been included at the end of the manuscript. In addition, all in-text citations have been carefully revised to ensure full consistency with the corresponding Supporting Information labels.

Furthermore, to improve clarity and conciseness of the manuscript, several references (Refs. 8, 9, 11, 13, 14, 15, 16, 17, 25, and 27) have been removed from the Introduction and Discussion sections. Accordingly, the reference list has been updated and renumbered, and all in-text citations have been revised to ensure consistency throughout the manuscript.

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

Reviewer #1: Partly

Reviewer #2: Yes

Response to Reviewer: The manuscript has been carefully revised to further strengthen its technical robustness and ensure that all conclusions are fully supported by the data presented.

Specifically, the following improvements have been made:

• The experimental design, including controls, replication, and sampling procedures, has been clarified in the Materials and Methods section to ensure transparency and reproducibility.

• Statistical analyses have been revised and reported consistently in accordance with journal guidelines (e.g., F-values, degrees of freedom, and p-values), ensuring clarity and accuracy.

• Seasonal mean abundances (± SEM) have been calculated and presented in a newly structured table to allow clearer comparison among treatments, as recommended.

• Data presentation has been simplified, with detailed temporal trends moved to supplementary materials, improving readability and interpretation of results.

• The Results and Discussion sections have been refined to ensure that all conclusions are directly supported by the data, avoiding overinterpretation.

These revisions have substantially improved the clarity, rigor, and consistency of the manuscript. Therefore, the study now fully meets the criteria for technical soundness, with conclusions appropriately supported by the presented data.

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: I Don't Know

Reviewer #2: Yes

Response to Reviewer: The statistical analysis has been carefully revised and clarified throughout the manuscript to ensure transparency and reproducibility.

To address potential ambiguity, the following improvements have been made:

• The statistical methods have been described in greater detail in the Statistical Analysis section, including the specific models used, assumptions tested, and post-hoc procedures applied.

• Repeated-measures ANOVA has been clearly specified as the primary analytical approach for evaluating treatment effects over time.

• All statistical results are now reported consistently in accordance with journal guidelines (e.g., F-values with exact p-values).

• Post-hoc comparisons (Tukey’s HSD) are explicitly indicated, and their application is clearly explained.

• Data presentation has been improved by including seasonal mean values (± SEM), allowing clearer interpretation of treatment effects.

These revisions ensure that the statistical analyses are both rigorous and clearly presented. The uncertainty expressed by Reviewer #1 is likely due to insufficient detail in the previous version, which has now been fully addressed.

3. Have the authors made all data underlying the findings in their manuscript fully available?

Reviewer #1: Yes

Reviewer #2: Yes

4. Is the manuscript presented in an intelligible fashion and written in standard English?

Reviewer #1: No

Reviewer #2: Yes

Response to Reviewer: The manuscript has been thoroughly revised to improve clarity, readability, and overall language quality.

5. Review Comments to the Author

Response to Reviewer:

Thank you for this valuable and constructive comment. The Introduction has been carefully revised and substantially shortened to improve focus and clarity. In particular, the section has been restructured to better highlight the research gap and to clearly state the objectives of the study, thereby strengthening the overall rationale.

To address this concern, the following revisions have been made:

• Lines 52–53 and 59–64 have been removed to eliminate redundant background information.

• The literature presented in Lines 84–89 has been revised to focus specifically on neonicotinoids in relation to clothianidin, and the references have been updated accordingly.

• Lines 103–148 have been removed to reduce excessive literature review and improve conciseness.

• The Introduction has been streamlined and reduced to 559 words.

As a result, the revised Introduction is now more concise, better aligned with the study objectives, and more clearly structured around the identified research gap.

Response to Reviewer: Thank you for this valuable suggestion. I agree that conducting similar studies across different fields would provide broader insight and increase environmental variability. However, the present study was intentionally conducted within the same field under uniform conditions to minimize environmental heterogeneity and ensure comparability among treatments. This approach allowed us to focus specifically on treatment effects. Moreover, previous multi-site field studies and meta-analyses have reported consistent patterns regarding the effects of neonicotinoid seed treatments on natural enemy communities across different geographic locations [Douglas and Tooker, 2016; Saeed and Razaq, 2016], suggesting that major outcome differences among fields may be limited.

Douglas MR, Tooker JF. Meta-analysis reveals that seed-applied neonicotinoids and pyrethroids have similar negative effects on abundance of arthropod natural enemies. PeerJ. 2016 Dec 7;4:e2776. doi: 10.7717/peerj.2776. PMID: 27957400; PMCID: PMC5147019.

Saeed R, Razaq M, Hardy IC. Impact of neonicotinoid seed treatment of cotton on the cotton leafhopper, Amrasca devastans (Hemiptera: Cicadellidae), and its natural enemies. Pest Manag Sci. 2016 Jun;72(6):1260-7. doi: 10.1002/ps.4146. Epub 2015 Oct 5. PMID: 26436945.

Response to Reviewer: Sweep net sampling was used to assess the abundance of mobile foliar-dwelling arthropods present in the plant canopy. For hemimetabolous groups, including Miridae, Cicadellidae, Nabidae and Anthocoridae, both nymphs and adults captured by sweep netting were included in the counts because these taxa undergo incomplete metamorphosis and their nymphal stages are active and mobile within the canopy. In contrast, for holometabolous groups such as Chrysopidae and Asilidae, only adult individuals were recorded, as their larval stages occur in different microhabitats (e.g., soil or concealed substrates) and are not effectively sampled using sweep netting (Line173-179).

Sampling was conducted for twelve consecutive weeks during the cotton growing seasons of 2021 and 2022 (June–September). During each sampling event, 50 sweeps were performed per plot along a straight transect through the center of each plot. The experiment consisted of three replicates arranged in a randomized complete block design. Sweep netting was carried out at least 100 m from the field edge to minimize potential edge effects. The same standardized sampling protocol (50 sweeps per replicate per sampling date) was consistently applied across both years and treatments.

Response to Reviewer: Seasonal mean abundances (± SEM) for Mirida

Attachment

Submitted filename: MYResponse to the reviewers comments.docx

pone.0346422.s018.docx^{(63.7KB, docx)}

PLoS One. doi: 10.1371/journal.pone.0346422.r003

Decision Letter 1

Rachid Bouharroud

19 Mar 2026

Trade-offs in cotton pest management: Seed treatments suppress pests but reduce the abundance of natural enemies in the arthropod community.

PONE-D-25-50468R1

Dear Dr. Yalçın,

We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact and clicking the ‘Update My Information' link at the top of the page. For questions related to billing, please contact billing support....

If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

Kind regards,

Rachid Bouharroud

Academic Editor

PLOS One

Additional Editor Comments (optional):

Reviewers' comments:

PLoS One. doi: 10.1371/journal.pone.0346422.r004

Acceptance letter

Rachid Bouharroud

PONE-D-25-50468R1

PLOS One

Dear Dr. Yalçın,

I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS One. Congratulations! Your manuscript is now being handed over to our production team.

At this stage, our production department will prepare your paper for publication. This includes ensuring the following:

* All references, tables, and figures are properly cited

* All relevant supporting information is included in the manuscript submission,

* There are no issues that prevent the paper from being properly typeset

You will receive further instructions from the production team, including instructions on how to review your proof when it is ready. Please keep in mind that we are working through a large volume of accepted articles, so please give us a few days to review your paper and let you know the next and final steps.

Lastly, if your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

You will receive an invoice from PLOS for your publication fee after your manuscript has reached the completed accept phase. If you receive an email requesting payment before acceptance or for any other service, this may be a phishing scheme. Learn how to identify phishing emails and protect your accounts at https://explore.plos.org/phishing.

If we can help with anything else, please email us at customercare@plos.org.

Thank you for submitting your work to PLOS ONE and supporting open access.

Kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Rachid Bouharroud

Academic Editor

PLOS One

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

S1 Table. Mean sums of Cicadellidae and Miridae populations collected per plot using sweep nets.

(PDF)

pone.0346422.s001.pdf^{(149.5KB, pdf)}

(PDF)

pone.0346422.s002.pdf^{(198KB, pdf)}

S1 Fig. Impact of pesticide treatments on Cicadellidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s003.pdf^{(75KB, pdf)}

S2 Fig. Impact of pesticide treatments on Cicadellidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s004.pdf^{(76.4KB, pdf)}

S3 Fig. Impact of pesticide treatments on Miridae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s005.pdf^{(73.3KB, pdf)}

S4 Fig. Impact of pesticide treatments on Miridae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s006.pdf^{(71.3KB, pdf)}

S5 Fig. Impact of pesticide treatments on Anthocoridae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s007.pdf^{(94.7KB, pdf)}

S6 Fig. Impact of pesticide treatments on Anthocoridae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s008.pdf^{(103.2KB, pdf)}

S7 Fig. Impact of pesticide treatments on Asillidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s009.pdf^{(93.4KB, pdf)}

S8 Fig. Impact of pesticide treatments on Asillidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s010.pdf^{(118.1KB, pdf)}

S9 Fig. Impact of pesticide treatments on Coccinellidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s011.pdf^{(113.2KB, pdf)}

S10 Fig. Impact of pesticide treatments on Coccinellidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s012.pdf^{(113KB, pdf)}

S11 Fig. Impact of pesticide treatments on Chrysopidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s013.pdf^{(110.5KB, pdf)}

S12 Fig. Impact of pesticide treatments on Chrysopidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s014.pdf^{(98.7KB, pdf)}

S13 Fig. Impact of pesticide treatments on Nabidae abundance during crop development stages in 2021.

(PDF)

pone.0346422.s015.pdf^{(110.7KB, pdf)}

S14 Fig. Impact of pesticide treatments on Nabidae abundance during crop development stages in 2022.

(PDF)

pone.0346422.s016.pdf^{(95.2KB, pdf)}

Attachment

Submitted filename: MYResponse to the reviewers comments.docx

pone.0346422.s018.docx^{(63.7KB, docx)}

Data Availability Statement

All relevant data are within the manuscript and its Supporting Information files.

[pone.0346422.ref001] 1.Ünay A, Balcï S, Cïnar VM. Gene action and combining ability for neps and seed coat neps in cotton (Gossypium hirsutum L.). Turkish J Field Crops. 2019;24(2):211–4. doi: 10.17557/tjfc.568326 [DOI] [Google Scholar]

[pone.0346422.ref002] 2.Jabran K, Chauhan BS. Cotton production. Hoboken (NJ): John Wiley & Sons; 2019. doi: 10.1002/9781119385523 [DOI] [Google Scholar]

[pone.0346422.ref003] 3.Naveed M, Maqsood MF, Cheema AR. Modeling the contribution of district-level cotton production to aggregate cotton production in Punjab (Pakistan): an empirical evidence using correlated component regression approach. J Excell Soc Sci. 2024;3(3):147–64. doi: 10.69565/jess.v3i3.200 [DOI] [Google Scholar]

[pone.0346422.ref004] 4.Mollaee M, Mobli A, Mutti NK, Manalil S, Chauhan BS. Challenges and opportunities in cotton production. cotton production. Wiley; 2019. 371–90. doi: 10.1002/9781119385523.ch18 [DOI] [Google Scholar]

[pone.0346422.ref005] 5.Douglas MR, Tooker JF. Meta-analysis reveals that seed-applied neonicotinoids and pyrethroids have similar negative effects on abundance of arthropod natural enemies. PeerJ. 2016;4:e2776. doi: 10.7717/peerj.2776 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref006] 6.Smalling KL, Hladik ML, Sanders CJ, Kuivila KM. Leaching and sorption of neonicotinoid insecticides and fungicides from seed coatings. J Environ Sci Health B. 2018;53(3):176–83. doi: 10.1080/03601234.2017.1405619 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref007] 7.Bandeira FO, Alves PRL, Hennig TB, Brancalione J, Nogueira DJ, Matias WG. Chronic effects of clothianidin to non-target soil invertebrates: ecological risk assessment using the species sensitivity distribution (SSD) approach. J Hazard Mater. 2021;419:126491. doi: 10.1016/j.jhazmat.2021.126491 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref008] 8.Hilton MJ, Emburey SN, Edwards PA, Dougan C, Ricketts DC. The route and rate of thiamethoxam soil degradation in laboratory and outdoor incubated tests, and field studies following seed treatments or spray application. Pest Manag Sci. 2019;75(1):63–78. doi: 10.1002/ps.5168 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref009] 9.Bartlett DW, Clough JM, Godwin JR, Hall AA, Hamer M, Parr-Dobrzanski B. The strobilurin fungicides. Pest Manag Sci. 2002;58(7):649–62. doi: 10.1002/ps.520 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref010] 10.Moser SE, Obrycki JJ. Non-target effects of neonicotinoid seed treatments; mortality of coccinellid larvae related to zoophytophagy. Biological Control. 2009;51(3):487–92. doi: 10.1016/j.biocontrol.2009.09.001 [DOI] [Google Scholar]

[pone.0346422.ref011] 11.Gontijo PC, Moscardini VF, Michaud JP, Carvalho GA. Non-target effects of two sunflower seed treatments on Orius insidiosus (Hemiptera: Anthocoridae). Pest Manag Sci. 2015;71(4):515–22. doi: 10.1002/ps.3798 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref012] 12.Özşirvan G, Yalçın M, Turgut N, Tari V, Turgut C. Insights into the uptake, translocation, and accumulation dynamics of cyantraniliprole and thiamethoxam seed coating pesticides in maize plants. Environ Sci Pollut Res Int. 2024;31(32):44900–7. doi: 10.1007/s11356-024-34135-7 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref013] 13.Rein A, Trapp S, Fantke P, Yalçın M, Turgut N, Ahat C, et al. Uptake and translocation of pesticides in pepper and tomato plants. Pest Manag Sci. 2025;81(3):1562–70. doi: 10.1002/ps.8556 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref014] 14.Dubey A, Lewis MT, Dively GP, Hamby KA. Ecological impacts of pesticide seed treatments on arthropod communities in a grain crop rotation. J Appl Ecol. 2020;57(5):936–51. doi: 10.1111/1365-2664.13595 [DOI] [Google Scholar]

[pone.0346422.ref015] 15.Yalçin M, Kapiz I, Kösoglu M, Koca YO, Turgut N, Topal E, et al. Translocation of clothianidin to guttation fluid and its potential impact on honey bee, Apis mellifera anatoliaca Maa, 1953 (Hymenoptera: Apidae). Turk Entomol Derg. 2021;45(4):511–22. doi: 10.16970/entoted.1017479 [DOI] [Google Scholar]

[pone.0346422.ref016] 16.Moscardini VF, Gontijo PC, Michaud JP, Carvalho GA. Sublethal effects of insecticide seed treatments on two nearctic lady beetles (Coleoptera: Coccinellidae). Ecotoxicology. 2015;24(5):1152–61. doi: 10.1007/s10646-015-1462-4 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref017] 17.Seagraves MP, Lundgren JG. Effects of neonicitinoid seed treatments on soybean aphid and its natural enemies. J Pest Sci. 2011;85(1):125–32. doi: 10.1007/s10340-011-0374-1 [DOI] [Google Scholar]

[pone.0346422.ref018] 18.Bredeson MM, Lundgren JG. Thiamethoxam seed treatments have no impact on pest numbers or yield in cultivated sunflowers. J Econ Entomol. 2015;108(6):2665–71. doi: 10.1093/jee/tov249 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref019] 19.Jones AG, Hoover K, Pearsons K, Tooker JF, Felton GW. Potential impacts of translocation of neonicotinoid insecticides to cotton (Gossypium hirsutum (Malvales: Malvaceae)) extrafloral nectar on parasitoids. Environ Entomol. 2020;49(1):159–68. doi: 10.1093/ee/nvz157 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref020] 20.Szczepaniec A. Interactive effects of crop variety, insecticide seed treatment, and planting date on population dynamics of sugarcane aphid (Melanaphis sacchari) and their predators in late-colonized sorghum. Crop Protection. 2018;109:72–9. doi: 10.1016/j.cropro.2018.03.002 [DOI] [Google Scholar]

[pone.0346422.ref021] 21.Pemberton RW, Lee JH. The influence of extrafloral nectaries on parasitism of an insect herbivore. Am J Bot. 1996;83(9):1187–94. doi: 10.1002/j.1537-2197.1996.tb13900.x [DOI] [Google Scholar]

[pone.0346422.ref022] 22.Nicolson SW, Massimo N, Pacini E. Nectaries and nectar. Nicolson SW, Nepi M, Pacini E, editors. 2007. doi: 10.1007/978-1-4020-5937-7 [DOI] [Google Scholar]

[pone.0346422.ref023] 23.Gao A-H, Fu Y-Y, Zhang K-Z, Zhang M, Jiang H-W, Fan L-X, et al. Azoxystrobin, a mitochondrial complex III Qo site inhibitor, exerts beneficial metabolic effects in vivo and in vitro. Biochim Biophys Acta. 2014;1840(7):2212–21. doi: 10.1016/j.bbagen.2014.04.002 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref024] 24.Smith JF, Catchot AL, Musser FR, Gore J. Effects of aldicarb and neonicotinoid seed treatments on twospotted spider mite on cotton. J Econ Entomol. 2013;106(2):807–15. doi: 10.1603/ec10125 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref025] 25.Disque HH, Hamby KA, Dubey A, Taylor C, Dively GP. Effects of clothianidin-treated seed on the arthropod community in a mid-Atlantic no-till corn agroecosystem. Pest Manag Sci. 2019;75(4):969–78. doi: 10.1002/ps.5201 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref026] 26.Shannon CE. A Mathematical theory of communication. Bell System Tech J. 1948;27(3):379–423. doi: 10.1002/j.1538-7305.1948.tb01338.x [DOI] [Google Scholar]

[pone.0346422.ref027] 27.Firake DM, Behere GT. Natural mortality of invasive fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) in maize agroecosystems of northeast India. Bio Control. 2020;148:104303. doi: 10.1016/j.biocontrol.2020.104303 [DOI] [Google Scholar]

[pone.0346422.ref028] 28.Oksanen J, Blanchet FG, Minchin PR. Vegan: community ecology package. [Google Scholar]

[pone.0346422.ref029] 29.Alberdi A, Gilbert MTP. Hilldiv: an R package for the integral analysis of diversity based on Hill numbers. 2019. http://biorxiv.org/lookup/doi/10.1101/545665

[pone.0346422.ref030] 30.Saeed R, Razaq M, Hardy IC. Impact of neonicotinoid seed treatment of cotton on the cotton leafhopper, Amrasca devastans (Hemiptera: Cicadellidae), and its natural enemies. Pest Manag Sci. 2016;72(6):1260–7. doi: 10.1002/ps.4146 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref031] 31.Mourtzinis S, Krupke CH, Esker PD, Varenhorst A, Arneson NJ, Bradley CA, et al. Neonicotinoid seed treatments of soybean provide negligible benefits to US farmers. Sci Rep. 2019;9(1):11207. doi: 10.1038/s41598-019-47442-8 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref032] 32.Regan K, Ordosch D, Glover KD, Tilmon KJ, Szczepaniec A. Effects of a pyrethroid and two neonicotinoid insecticides on population dynamics of key pests of soybean and abundance of their natural enemies. Crop Protection. 2017;98:24–32. doi: 10.1016/j.cropro.2017.03.004 [DOI] [Google Scholar]

[pone.0346422.ref033] 33.Özgür O, Atakan E, Pehlïvan S. Investigation of the damage of Miridae species on cotton in Çukurova Region of Turkey. Turk Entomol Derg. 2019;43(2):143–56. doi: 10.16970/entoted.517428 [DOI] [Google Scholar]

[pone.0346422.ref034] 34.Lu Y, Wu K, Jiang Y, Xia B, Li P, Feng H, et al. Mirid bug outbreaks in multiple crops correlated with wide-scale adoption of Bt cotton in China. Science. 2010;328(5982):1151–4. doi: 10.1126/science.1187881 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref035] 35.Jiao Z, Jaworski CC, Lu Y, Ye L, Wu K, Desneux N. Maize fields are a potential sink for an outbreaking mirid bug pest in Chinese Bt-cotton agricultural landscapes. Agric Ecosyst Environ. 2019;279:122–9. doi: 10.1016/j.agee.2019.04.010 [DOI] [Google Scholar]

[pone.0346422.ref036] 36.Zhang Z, Wang Y, Zhao Y, Li B, Lin J, Zhang X, et al. Nitenpyram seed treatment effectively controls against the mirid bug Apolygus lucorum in cotton seedlings. Sci Rep. 2017;7(1):8573. doi: 10.1038/s41598-017-09251-9 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref037] 37.Delbeke E, Vercruysse P, Tirry L, De Clercq P, Degheele D. Toxicity of diflubenzuron, pyriproxyfen, imidacloprid and diafenthiuron to the predatory bug Orius laevigatus (Het: Anthacoridae). Entomophaga. 1997;42(3):349–58. doi: 10.1007/BF02769828 [DOI] [Google Scholar]

[pone.0346422.ref038] 38.Desneux N, Decourtye A, Delpuech J-M. The sublethal effects of pesticides on beneficial arthropods. Annu Rev Entomol. 2007;52:81–106. doi: 10.1146/annurev.ento.52.110405.091440 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref039] 39.Papachristos DP, Milonas PG. Adverse effects of soil applied insecticides on the predatory coccinellid Hippodamia undecimnotata (Coleoptera: Coccinellidae). Biological Control. 2008;47(1):77–81. doi: 10.1016/j.biocontrol.2008.06.009 [DOI] [Google Scholar]

[pone.0346422.ref040] 40.Peck DC. Long-term effects of imidacloprid on the abundance of surface- and soil-active nontarget fauna in turf. Agric For Entomol. 2009;11(4):405–19. doi: 10.1111/j.1461-9563.2009.00454.x [DOI] [Google Scholar]

[pone.0346422.ref041] 41.Yao F-L, Zheng Y, Zhao J-W, Desneux N, He Y-X, Weng Q-Y. Lethal and sublethal effects of thiamethoxam on the whitefly predator Serangium japonicum (Coleoptera: Coccinellidae) through different exposure routes. Chemosphere. 2015;128:49–55. doi: 10.1016/j.chemosphere.2015.01.010 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref042] 42.Tappert L, Pokorny T, Hofferberth J, Ruther J. Sublethal doses of imidacloprid disrupt sexual communication and host finding in a parasitoid wasp. Sci Rep. 2017;7:42756. doi: 10.1038/srep42756 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref043] 43.Albajes R, López C, Pons X. Predatory fauna in cornfields and response to imidacloprid seed treatment. J Econ Entomol. 2003;96(6):1805–13. doi: 10.1093/jee/96.6.1805 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref044] 44.Pisa LW, Amaral-Rogers V, Belzunces LP, Bonmatin JM, Downs CA, Goulson D, et al. Effects of neonicotinoids and fipronil on non-target invertebrates. Environ Sci Pollut Res Int. 2015;22(1):68–102. doi: 10.1007/s11356-014-3471-x [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref045] 45.Feese H, Wilde G. Planting time applications of systemic insecticides on grain sorghum for greenbug control: Interactions with herbicides and effect on predators (Hemiptera (Homoptera)). J Kans Entomol Soc. 1975;48(3):396–402. [Google Scholar]

[pone.0346422.ref046] 46.Al-Deeb MA, Wilde GE, Zhu KY. Effect of insecticides used in corn, sorghum, and alfalfa on the predator Orius insidiosus (Hemiptera: Anthocoridae). J Econ Entomol. 2001;94(6):1353–60. doi: 10.1603/0022-0493-94.6.1353 [DOI] [PubMed] [Google Scholar]

[pone.0346422.ref047] 47.Lundgren JG, Fausti SW. Trading biodiversity for pest problems. Sci Adv. 2015;1(6):e1500558. doi: 10.1126/sciadv.1500558 [DOI] [PMC free article] [PubMed] [Google Scholar]

[pone.0346422.ref048] 48.Schneider CW, Tautz J, Grünewald B, Fuchs S. RFID tracking of sublethal effects of two neonicotinoid insecticides on the foraging behavior of Apis mellifera. PLoS One. 2012;7(1):e30023. doi: 10.1371/journal.pone.0030023 [DOI] [PMC free article] [PubMed] [Google Scholar]

PERMALINK

Trade-offs in cotton pest management: Seed treatments suppress pests but reduce the abundance of natural enemies in the arthropod community

Melis Yalçin

Roles

Abstract

Introduction

Materials and methods

Study site

Experimental setup and arthropod collection methods

Statistical analysis

Results

Effects on pest communities

Table 1. Seasonal mean abundance of Miridae and Cicadellidae under different seed treatments in 2021 and 2022.

Effects on predator communities

Table 2. Mean sums of cotton natural enemy population collected per plot using sweep nets.

Community-level implications

Fig 1. Principle response curve (PRC) exhibiting the effects of different seed treatments on arthropod communities during the vegetation period of cotton.

Table 3. Results (F-values) of repeated-measures (weeks) ANOVA of the effects of seed treatments, years, and their interaction on the Shannon-Wiener and Evenness Index of predators.

Table 4. The comparison of seed treatments regarding Shannon-Wiener and Evenness Index in each year separately.

Discussion

Effects of pesticides on pest communities

Effects of pesticides on predator communities

Effects of pesticides on arthropod communities

Conclusion

Supporting information

Data Availability

Funding Statement

References

Decision Letter 0

Rachid Bouharroud

Roles

Author response to Decision Letter 1

Decision Letter 1

Rachid Bouharroud

Roles

Acceptance letter

Rachid Bouharroud

Roles

Associated Data

Supplementary Materials

Data Availability Statement

ACTIONS

PERMALINK

RESOURCES

Similar articles

Cited by other articles

Links to NCBI Databases